Isolation of the Gene Encoding the Hin Recombinational Enhancer Binding Protein

Creators: Johnson, Reid C.; Ball, Catherine A.; Pfeffer, Diana; Simon, Melvin I.

Abstract

In vitro DNA inversion mediated by the protein Hin requires the presence of a recombinational enhancer sequence located in cis relative to the recombination sites and a protein, Fis, which binds to the enhancer. We have cloned and determined the primary sequence of the gene encoding Fis. The deduced amino acid sequence of Fis indicates that the protein is 98 amino acids long and contains a potential helix-turn-helix DNA binding motif at its carboxyl terminus. The gene encoding Fis maps at 72 min on the Escherichia coli chromosome. The construction of mutant strains of E. coli that lack a functional fis gene demonstrates that Fis is not essential for cell growth under laboratory conditions but is required for high rates of Hin-mediated site-specific inversion in vivo.

Additional Information

© 1988 by the National Academy of Sciences Contributed by Melvin I. Simon, December 23, 1987. Note Added in Proof. R. Weisberg (National Institutes of Health) has pointed out that the NtrC-like protein from "Bradyrhizobium parasponiae" (36) displays greater homology to the E. coli Fis protein than does the K. pneumoniae NtrC. In the carboxyl-terminal region of the two proteins, there are 16 identities over a 22-amino acid region, and statistically significant homology extends over the entire Fis sequence. We thank B. Bachmann and C. Gross for strains, D. Glitz and S. Horvath for oligonucleotide synthesis, L. Williams for protein sequencing, and A. Glasgow and K. Hughes for critically reading the manuscript. This work was supported by a grant from the California Institute for Cancer Research, a Basil O'Connor Starter Scholar Research Award No. 5-623 from the March of Dimes Birth Defects Foundation, the Searle Scholars Program/The Chicago Community Trust, and grant GM38509 from the National Institutes of Health to R.C.J. C.A.B. was supported in part by National Research Service Award GM07104 from the National Institutes of Health. M.I.S. was supported by a grant from the National Science Foundation. The sequence reported in this paper is being deposited in the EMBL/GenBank data base (Bolt, Beranek, and Newman Laboratories, Cambridge, MA, and Eur. Mol. Biol. Lab., Heidelberg) (accession no. J03245). The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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